LOCUS       sequence-346860-        5534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Chlamydomonas reinhardtii RSP10recombinant protein in 
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5534)
  AUTHORS   Grossman-Haham I, Coudray N, Yu Z, Wang F, Zhang N, Bhabha G, Vale 
            RD
  TITLE     Structure of the radial spoke head and insights into its role in 
            mechanoregulation of ciliary beating.
  JOURNAL   Nat Struct Mol Biol. 2021 Jan;28(1):20-28. doi: 
            10.1038/s41594-020-00519-9. Epub 2020 Dec 14.
  PUBMED    33318704
REFERENCE   2  (bases 1 to 5534)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41594-020-00519-9"; journalName: "Nat Struct Mol Biol"; 
            date: "2021-01"; volume: "28"; issue: "1"; pages: "20-28"
COMMENT     Sequence Label: pLIB-RSP10
FEATURES             Location/Qualifiers
     source          1..5534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(115..120,121..137,138..143))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(539..572)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        620..711
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     636..653
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    1534..1668
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1571..1590)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1625..1644
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1778..1943
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2127..2581
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2214..2233)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2423..2444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2607..2711
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2712..2780,2781..3572)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2930..2949)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3743..4331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4232..4251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4485..4502
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4636..4860
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4927..5460)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 tgtacaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg
       61 cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca ttacagttta
      121 cgaaccgaac aggcttatgt caactgggtt tgtgccttca tccgtttcca cggtgtgcgt
      181 cacccggcaa ccttgggcag cagcgaagtc gaggcatttc tgtcctggct ggcgaacgag
      241 cgcaaggttt cggtctccac gcatcgtcag gcattggcgg ccttgctgtt cttctacggc
      301 aaggtgctgt gcacggatct gcccttgctt caggagatcg gtagacctcg gccgtcgcgg
      361 cgcttgccgg tggtgctgac cccggatgaa gtggttcgca tcctcggttt tctggaaggc
      421 gagcatcgtt tgttcgccca ggactctagc tatagttcta gtggttggct acagctttgt
      481 ttgtactatc aacaggttga actgctgatc aacagatcct ctacgcggcc gcggtaccat
      541 aacttcgtat agcatacatt atacgaagtt atctggtttt actagtatcg attcgcgacc
      601 tactccggaa tattaataga tcatggagat aattaaaatg ataaccatct cgcaaataaa
      661 taagtatttt actgttttcg taacagtttt gtaataaaaa aacctataaa tattccggat
      721 tattcatacc gtcccaccat cgggcgcgga tccatggccg atgacgagct ccccccgcaa
      781 ccggtttggg agggtccact cgatgaagac ggtaaaccac atggactcgg caaaatggag
      841 tatcctccac cccctatggg cgaggatgat gaggaagaga agcccggtga caaattcgag
      901 ggaacgatgg agcatggagt tagaacggga aaaggcactt atacgtgggg agtgtccggc
      961 gctgtatata cgggagacta tgtgaatggc aaaaaacatg gcaaaggaaa aatggtttac
     1021 cctgacaaag gtgtctacga gggcgattgg gtcgaggatg taatgcaagg tcaaggtaca
     1081 tacacttatc ccaatggaga tatataccaa ggtgcttttt gggccggtaa gaggcacgga
     1141 aaaggtatgt accactataa aggaccctgt tgtcagttgg taggtgattg ggctgacggc
     1201 ggttttactt atggtcgttg ggtgtatgcc gacggttcca tgtttatggg aaaatttgga
     1261 ggagctgccg ccgacagcaa accaactgcg ggaagctact tttattccag ttcctcgctg
     1321 gtgcaagagg gacactttgc gaaggacggc agttgggtgg gccatcgcga ccctgcagtg
     1381 ggcaaggaat tttctgtcgc gtaaaagctt gtcgagaagt actagaggat cataatcagc
     1441 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1501 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1561 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1621 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt
     1681 gagcctagaa gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     1741 cgctgagcaa taactatcat aacccctagg agatccgaac cagataagtg aaatctagtt
     1801 ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac ccagttccca
     1861 tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc cctcccagtt
     1921 cccaactatt ttgtccgccc acagcggggc atttttcttc ctgttatgtt tttaatcaaa
     1981 catcctgcca actccatgtg acaaaccgtc atcttcggct actttttctc tgtcacagaa
     2041 tgaaaatttt tctgtcatct cttcgttatt aatgtttgta attgactgaa tatcaacgct
     2101 tatttgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     2161 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2221 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2281 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2341 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2401 gttggagtcc acgttcttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2461 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2521 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     2581 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2641 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2701 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2761 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2821 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2881 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2941 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3001 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3061 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3121 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3181 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3241 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3301 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3361 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3421 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3481 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3541 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3601 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3661 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3721 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3781 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3841 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactatt cttctagtgt
     3901 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3961 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4021 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4081 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4141 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4201 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4261 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4321 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4381 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4441 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4501 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4561 accgcataga ccagccgcgt aacctggcaa aatcggttac ggttgagtaa taaatggatg
     4621 ccctgcgtaa gcgggtgtgg gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa
     4681 ctatgacaat aaagtcttaa actagacaga atagttgtaa actgaaatca gtccagttat
     4741 gctgtgaaaa agcatactgg acttttgtta tggctaaagc aaactcttca ttttctgaag
     4801 tgcaaattgc ccgtcgtatt aaagaggggc gtggccaagg gcatggtaaa gactatattc
     4861 gcggcgttgt gacaatttac cgaacaactc cgcggccggg aagccgatct cggcttgaac
     4921 gaattgttag gtggcggtac ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc
     4981 caactttgta tagagagcca ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac
     5041 ataagcacca agcgcgttgg cctcatgctt gaggagattg atgagcgcgg tggcaatgcc
     5101 ctgcctccgg tgctcgccgg agactgcgag atcatagata tagatctcac tacgcggctg
     5161 ctcaaacttg ggcagaacgt aagccgcgag agcgccaaca accgcttctt ggtcgaaggc
     5221 agcaagcgcg atgaatgtct tactacggag caagttcccg aggtaatcgg agtccggctg
     5281 atgttgggag taggtggcta cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg
     5341 catggatttg acttggtcag ggccgagcct acatgtgcga atgatgccca tacttgagcc
     5401 acctaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat
     5461 cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg
     5521 cccgaggcat agac
//
